T cells harboring anti-CD20 scFv-CD8-TCRζ fusion gene in treatment of human B-cell lymphomas: An experimental sudy
- Author:
Lin YUN
1
Author Information
1. Department of Internal Medicine
- Publication Type:Journal Article
- From:
Academic Journal of Second Military Medical University
2006;27(7):745-749
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To observe the anti-tumor effect of T lymphocytes harboring anti-CD20 scFv-CD8-TCRζ fusion gene on human B lymphomas in vitro and in vivo, so as to explore the feasibility of CD20-mediated autogenous T lymphocytes in killing B-cell lymphomas. Methods: A fusion gene containing anti-CD20 scFv, CD8 molecule and CD3ζ chain was constructed and was cloned into pcDNA3. After confirmed by restriction endonuclease analysis, the fusion gene was used to transfect the human peripheral T lymphocytes through electroporation and expression of anti-CD20 scFv-CD8-TCRζ fusion protein was induced. The CD20 antigen-recognition ability of transfected T cells was determined by flow cytometry. The killing effect of transfected T cells on B cell lymphomas-Raji cells was tested in a cytotoxicity assay. The anti-tumor efficacy of this gene-modified T cell against the Raji tumor cell line was also evaluated in BALB/c nude mice. Results: We successfully constructed the anti-CD20 scFv-CD8-TCRζ fusion gene and expressed its protein on T cells. Flow cytometry and cytotoxicity assay demonstrated that the gene engineered T cells specifically recognized CD20 antigen and specifically inhibited B-cell lymphomas Raji cells. Furthermore, the T cells significantly inhibited the transplanted Raji cells in irradiated BALB/c nude mice. Conclusion: T lymphocytes transfected with anti-CD20 scFv-CD8-TCRζ fusion gene have antigen-specific anti-tumor effect in vitro and in vivo, which lays a foundation for utilizing human T lymphocytes to treat human B-cell lymphomas.
